Table 2 Plasmids used in this study

pZS4Int-tetR

Gene encoding the TetR repressor under control of the constitutive P_N25 promoter, SC101 ori, Sp^r

[45]

pMA7CR_2.0

λ Red β-protein under control of the L-arabinose inducible P_BAD, Cas9 under control of the anhydrotetracycline inducible P_Ltet, ColE1 ori, Amp^r

[30]

pMAZ-SK

sgRNA targeting galK under control of the anhydrotetracycline inducible P_Ltet, L-rhamnose inducible plasmid self-killing mechanism, ColA ori, Km^r

[30]

pMAZ-SK_x

pMAZ-SK with gRNA variants targeting x gene:

… dsbA

… dsbB

… skp

… sppA

… degP

This study

pFREE

Cas9 under control of the anhydrotetracycline inducible P_Ltet, gRNA targeting the most common replicons under control of the L-rhamnose inducible P_rha, ColA ori, tetR, Km^r

[31]

pSB-M1s

pelB-scFv173–2-5-phoA fusion gene under control of the m-toluate inducible XylS/Pm promoter system, RK2 replicon, Km^r

[25]

pGM29ompA

gene encoding GM-CSF with ompA signal sequence under control of the m-toluate inducible XylS/Pm promoter system, RK2 replicon, Amp^r

[26]